Targeting POLE2 Creates a Novel Vulnerability in Renal Cell Carcinoma via Modulating Stanniocalcin 1.
Targeting POLE2 Creates a Novel Vulnerability in Renal Cell Carcinoma via Modulating Stanniocalcin 1.
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DOI：
10.3389/fcell.2021.622344
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发表时间：
2021
影响因子：
5.5
通讯作者：
He H
中科院分区：
文献类型：
--
作者：
Zhang C;Shen Y;Gao L;Wang X;Huang D;Xie X;Xu D;He H
本研究旨在探讨DNA聚合酶α亚基2（POLE 2）在肾细胞癌（RCC）中的生物学功能及其作用机制。选择癌症基因组图谱肾透明细胞癌（TCGA-KIRC）和国际癌症基因组联盟（ICGC）数据库中的POLE 2表达数据集，分析POLE 2与各种临床病理参数的相关性。免疫组化法检测肾癌组织中POLE 2的表达。构建POLE 2敲低细胞系。本研究通过体外和体内实验研究了POLE 2在肾细胞癌细胞生物学中的作用，包括细胞存活率实验、克隆形成实验、流式细胞术、创伤愈合实验、Transwell实验、qRT-PCR、Western blot等，并采用基因芯片、免疫共沉淀、拯救实验和Western blot等方法研究了POLE 2的分子机制。POLE 2在肾细胞癌组织中呈高表达，POLE 2高表达与肾细胞癌预后不良相关。此外，敲低POLE 2在体外显著抑制细胞增殖、迁移并促进细胞凋亡。体内实验表明，POLE 2减弱RCC肿瘤发生和肿瘤生长。同时也说明斯钙素1（stanniocalcin 1，STC 1）是POLE 2的下游基因，在肾癌的发生发展中起重要作用。此外，POLE 2基因敲低可显著上调Bad和p21的表达水平，而HSP 70、IGF-I、IGF-II、survivin和sTNF-R1的表达水平则显著下调。Western印迹分析还显示，POLE 2的敲低抑制了癌症相关通路蛋白的表达水平，包括p-Akt、CCND 1、MAPK 9和PIK 3CA。POLE 2基因的敲除可通过调控STC 1抑制肾癌细胞的增殖和迁移，提示POLE 2-STC 1可能成为肾癌治疗的潜在靶点。
The aim of this study is to investigate the biological functions and the underlying mechanisms of DNA polymerase epsilon subunit 2 (POLE2) in renal cell carcinoma (RCC). The datasets of POLE2 expression in The Cancer Genome Atlas Kidney Clear Cell Carcinoma (TCGA-KIRC) and International Cancer Genome Consortium (ICGC) databases was selected and the correlation between POLE2 and various clinicopathological parameters was analyzed. The POLE2 expression in RCC tissues was examined by immunohistochemistry. The POLE2 knockdown cell lines were constructed. In vitro and in vivo experiments were carried out to investigate the function of POLE2 on cellular biology of RCC, including cell viability assay, clone formation assay, flow cytometry, wound-healing assay, Transwell assay, qRT-PCR, Western blot, etc. Besides, microarray, co-immunoprecipitation, rescue experiment, and Western blot were used to investigate the molecular mechanisms underlying the functions of POLE2. POLE2 was overexpressed in RCC tissues, and high expression of POLE2 was correlated with poor prognosis of RCC. Furthermore, knockdown of POLE2 significantly inhibited cell proliferation, migration, and facilitated apoptosis in vitro. In vivo experiments revealed that POLE2 attenuated RCC tumorigenesis and tumor growth. we also illuminated that stanniocalcin 1 (STC1) was a downstream gene of POLE2, which promoted the occurrence and development of RCC. Besides, knockdown of POLE2 significantly upregulated the expression levels of Bad and p21 while the expression levels of HSP70, IGF-I, IGF-II, survivin, and sTNF-R1 were significantly downregulated. Western blot analysis also showed that knockdown of POLE2 inhibited the expression levels of Cancer-related pathway proteins including p-Akt, CCND1, MAPK9, and PIK3CA. Knockdown of POLE2 attenuates RCC cells proliferation and migration by regulating STC1, suggesting that POLE2-STC1 may become a potential target for RCC therapy.
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影响因子：
14.9
作者：
Ritchie ME;Phipson B;Wu D;Hu Y;Law CW;Shi W;Smyth GK
通讯作者：
Smyth GK
影响因子：
7.4
作者：
Ma X;Gu L;Li H;Gao Y;Li X;Shen D;Gong H;Li S;Niu S;Zhang Y;Fan Y;Huang Q;Lyu X;Zhang X
通讯作者：
Zhang X
影响因子：
11.2
作者：
Kim, Eun Mi;Jung, Chan-Hun;Um, Hong-Duck
通讯作者：
Um, Hong-Duck
DOI：
10.1159/000086042
发表时间：
2005-01-01
期刊：
CELLULAR STRESS RESPONSES IN RENAL DISEASES
影响因子：
--
作者：
Atkins, D;Lichtenfels, R;Seliger, B
通讯作者：
Seliger, B
影响因子：
3.4
作者：
Xiong, Yan;Wang, Qibai
通讯作者：
Wang, Qibai